U.S. patent number 5,151,509 [Application Number 07/285,510] was granted by the patent office on 1992-09-29 for gene encoding serine protease inhibitor.
This patent grant is currently assigned to United States of America. Invention is credited to Girish J. Kotwal, Bernard Moss.
United States Patent |
5,151,509 |
Kotwal , et al. |
September 29, 1992 |
Gene encoding serine protease inhibitor
Abstract
Novel serine protease inhibitors and genes encoding the same are
described.
Inventors: |
Kotwal; Girish J. (Bethesda,
MD), Moss; Bernard (Bethesda, MD) |
Assignee: |
United States of America
(Washington, DC)
|
Family
ID: |
23094557 |
Appl.
No.: |
07/285,510 |
Filed: |
December 16, 1988 |
Current U.S.
Class: |
536/23.2;
435/69.2 |
Current CPC
Class: |
C07K
14/811 (20130101) |
Current International
Class: |
C07K
14/81 (20060101); C12N 015/15 () |
Field of
Search: |
;435/69.2,183,172.3,320.1 ;536/27 ;935/14,10,9 |
Other References
Boursnell et al. J Gen Virol. 69:2995 (1988). .
Pickup et al. PNAS, 83:7698 (1986). .
Niles et al. Virology 153(1):96 (1986). .
Kotwal et al., "Vaccinia Virus Encodes Two Proteins That Are
Structurally Related to Members of the Plasma Serine Protease
Inhibitor Superfamily", Journal of Virology, Feb. 1989, vol. 63 No.
2, pp. 600-606..
|
Primary Examiner: Schwartz; Richard A.
Assistant Examiner: LeGuyader; John
Attorney, Agent or Firm: Townsend and Townsend
Claims
What is claimed is:
1. A cloned gene having the nucleotide sequence of FIG. 2A.
2. A cloned gene having the nucleotide sequence of FIG. 2A and
deposited at the ATCC under accession number 67864.
Description
The present invention is related generally to the identification
and characterization of new genes and proteins. More particularly,
the present invention is related to the identification of genes
which encode proteins having substantial degree of homology to the
serine protease inhibitor superfamily. There are no known synthetic
or microbial proteins capable of specifically inhibiting serine
proteases.
SUMMARY OF THE INVENTION
It is, therefore, an object of the present invention to provide
isolated, substantially pure, novel serine protease inhibitors.
It is another object of the present invention to provide a
nucleotide sequence directing the synthesis of novel serine
protease inhibitor, when cloned in a suitable expression
vector.
It is a further object of the present invention to provide a
therapeutic composition and method for treating or controlling
conditions that would benefit from inhibition of serine proteases
or that result from the deficiency of serine protease inhibitors,
such as emphysema, cirrhosis, liver cancer and the like.
Other objects and advantages will become evident from the following
detailed description of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
These and other objects, features and many of the attendant
advantages of the invention will be better understood upon a
reading of the following detailed description when considered in
connection with the accompanying drawings wherein:
FIG. 1 shows Hind III map of the vaccinia WR strain with the
enlarged right end. The vertical bars indicate the inverted
terminal repeats. The arrows indicate the direction and the
position of the 2 genes, numbered as I and II with similarities to
the serine protease inhibitor family. The letters H and S indicate
the restriction sites Hind III and SalI, respectively.
FIGS. 2A (parts 1-3) and 2B show the nucleotide sequence of the
genes I and II, respectively, of vaccinia virus (WR strain) in
5'-3' direction. The predicted coding sequence starting with the
initiation codon ATG to the termination codon has been translated
into the corresponding amino acid sequence which is shown below or
above the nucleotide sequence in the single letter code. The
nucleotide sequence was determined by the standard dideoxy-chain
termination technique using denatured plasmids as templates and
specific oligonucleotides as primers.
FIGS. 3A (parts 1-16) and 3B (parts 1-14) present the alignment of
the deduced amino acid sequence of the genes I and II,
respectively, of vaccinia virus with some of the members of the
serine protease inhibitor superfamily. Members of the superfamily
with significant similarities include human plasma serine protease
(protein C) inhibitor, human placental plasminogen activator and
human beta-migrating plasminogen activator. Dissimilar amino acids
have been highlighted by asterisks (*). Gaps imposed to maximize
alignment are indicated by empty spaces. The similarity was found
by searching the National Biomedical Research Foundation (NBRF)
protein data bank using the Beckman Instruments microgenie
program.
DETAILED DESCRIPTION OF THE INVENTION
The above and various other objects and advantages of the present
invention are achieved by the constructions having the nucleotide
and amino acid sequences as shown in FIGS. 2A and 2B.
Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. Although
any methods and materials similar or equivalent to those described
herein can be used in the practice or testing of the present
invention, the preferred methods and materials are now described.
All publications mentioned hereunder are incorporated herein by
reference. Unless mentioned otherwise, the techniques employed
herein are standard methodologies well known to one of ordinary
skill in the art.
The term "substantial homology" as used herein means having
homology greater than 25%.
The term "substantially pure" as used herein means as pure as can
be obtained by standard purification techniques.
Cloning of the genes is achieved by standard recombinant genetic
methodologies well known to one of ordinary skill in the art.
Given the cloned genes and the amino acid sequence, the proteins
can be synthesized by such methods as commercially available
peptide synthesizers or by employing suitable expression vectors
and the like, all of such methodologies being well known to one of
ordinary skill in the art to which this invention belongs. (See for
example Maniatis et al, "Molecular Cloning, A Laboratory Manual,"
Cold Spring Harbor, N.Y.)
A therapeutic composition in accordance with the present invention
comprises effective amount of the serine protease inhibitor to
inhibit serine protease activity and a pharmaceutically acceptable
carrier, if necessary.
A method for treating or controlling a condition resulting from the
deficiency of serine protease inhibitor or that would benefit from
inhibition of a serine protease comprises administering to a
subject afflicted with said condition, an effective amount of the
serine protease inhibitor to alleviate the deficiency of said
inhibitor.
A deposit of the embodiments of the present invention has been made
at the ATCC on Dec. 16, 1988. The deposit shall be viably
maintained, replacing if it becomes non-viable, for a period of 30
years from the date of the deposit, or for 5 years from the last
date of request for a sample of the deposit, whichever is longer,
and made available to the public without restriction in accordance
with the provisions of the law. The Commissioner of Patents and
Trademarks, upon request, shall have access to the deposit
(accession numbers 67864 and 67865).
It is understood that the examples and embodiments described herein
are for illustrative purposes only and that various modifications
or changes in light thereof will be suggested to persons skilled in
the art and are to be included within the spirit and purview of
this application and scope of the appended claims.
* * * * *